A v(d)j amplification optimization kit for immune repertoire sequencing

Abstract

The utility model discloses a V (D) J amplification optimization kit for immune group library sequencing relates to the field of kit, the utility model discloses an enzyme labeled plate and enzyme labeled strip, the enzyme labeled plate includes the board frame, the inside of board frame is provided with the board groove, the inner wall of board groove top is provided with a plurality of for the location structure of carrying ear, through setting locking part, fixed hole and connecting hole, wherein the location structure of board groove top is like carrying groove and the shallow position carrying cooperation of the ear of both sides of enzyme labeled strip, avoids the plug -in force of too tight, solves the enzyme labeled strip of prior art and is difficult to take out, and the problem of easy fracture because of too tight, and through setting in the board frame top both sides, by the locking part such as locking piece of torsional spring rotation connection to complete final fixation, when needing to take out enzyme labeled strip, just loosen the bolt, and locking piece will be opened automatically under the action of torsional spring, and enzyme labeled strip is completely released, and the operator can easily take out it from the location structure vertically.

Description

Technical Field

[0001] This invention relates to the field of reagent kits, specifically a V(D)J amplification optimization kit for immune repertoire sequencing. Background Technology

[0002] The V(D)J Amplification Optimization Kit for Immunome Repertoire Sequencing is a kit specifically designed for efficient and specific amplification of the variable region of the immune receptor (i.e., the V(D)J region) mediated by gene recombinase. This kit significantly improves the amplification efficiency and accuracy of V(D)J region gene fragments through various factors, including fidelity DNA polymerase, specific enhancers, buffers, and amplification program parameters. It effectively reduces non-specific amplification and bias, thus providing high-quality, highly representative DNA templates for subsequent immunoome repertoire sequencing. This helps researchers deeply understand the diversity, specificity, and dynamic changes of the immune system, and has broad and important application value in multiple research fields such as tumor immunology, infection immunology, and autoimmune diseases.

[0003] However, in current V(D)J amplification optimization kits used for immune repertoire sequencing, the microplates and strips are tightly fitted together. This is because the strips are secured to the plate via a specific slot structure to ensure stability during reaction preparation and incubation, preventing movement or displacement that could affect the accuracy and reproducibility of experimental results. However, the strips themselves are thin and flexible, and the locking structure is designed to be relatively tight for stability. This means that operators need to apply external force when removing them. If the operator is not skilled enough or uses incorrect techniques (e.g., applying the wrong direction, too much force, or uneven force), the strips can easily break. A broken strip not only spills the added sample or reagents, causing sample loss and reagent waste, but also contaminates the work surface and other equipment, increasing the risk of cross-contamination. Utility Model Content

[0004] Based on this, the purpose of this utility model is to provide a V(D)J amplification optimization kit for immune repertoire sequencing, in order to solve the technical problem that the enzyme strips in the existing kits are easily broken when they are removed due to their tight fit with the enzyme plate and their thin and flexible material, which leads to sample or reagent spillage, sample and reagent waste, and increased risk of cross-contamination.

[0005] To achieve the above objectives, this utility model provides the following technical solution: a V(D)J amplification optimization kit for immune repertoire sequencing, comprising an enzyme-labeled plate and enzyme-labeled strips, wherein the enzyme-labeled plate comprises a plate frame, the inner side of the plate frame is provided with a plate groove for holding the enzyme-labeled strips, the upper part of the plate groove is provided with a plurality of positioning structures for positioning the enzyme-labeled strips, and the top two sides of the plate frame are provided with side ears, and the side ears are rotatably connected by torsion springs to locking components for pressing and fixing the enzyme-labeled strips; The locking component has a connecting hole, and the top of the plate frame has a fixing hole corresponding to the connecting hole. When the locking component is flipped to the pressed state, the connecting hole and the fixing hole are aligned and locked by fasteners.

[0006] By adopting the above technical solution and combining the enzyme-labeled plate and enzyme-labeled strip, a stable and suitable carrier is provided for the V(D)J amplification experiment of immune repertoire sequencing, ensuring the orderly conduct of experimental operations and helping to improve experimental efficiency and the reliability of results.

[0007] Furthermore, the enzyme label strip includes multiple drop tubes, which are fixedly connected to each other, and two mounting ears are respectively provided on both sides of the row of drop tubes; the positioning structure is a mounting groove opened on the upper part of the inner wall of the plate groove, and the mounting ears are mounted with the mounting groove.

[0008] By adopting the above technical solution, the combination of the dropper on the enzyme-labeled strip and the mounting ear facilitates sample addition and fixation on the enzyme-labeled plate, making experimental operations more convenient, reducing the risk of sample spillage, and ensuring the smooth conduct of experiments.

[0009] Furthermore, the inner wall of the plate groove is provided with multiple insertion holes for inserting multiple sets of enzyme label strips in a row.

[0010] By adopting the above technical solution, the insertion holes at the bottom of the inner wall of the plate slot can further stabilize the insertion of multiple sets of enzyme label strips, prevent them from shaking and shifting during the experiment, ensure the stability of the experimental system, and improve the accuracy of the experiment.

[0011] Furthermore, the dropper has a flat bottom design, and the pore walls are surface-treated to enhance protein binding ability.

[0012] By adopting the above technical solution, the flat-bottom design of the dropper and the surface treatment of the orifice wall can enhance the protein binding capacity, facilitate the full reaction of the sample and reagent, improve the reaction efficiency, and make the experimental results more accurate and reliable.

[0013] Furthermore, the mounting ear and positioning structure are unidirectional embedding structures with matching shapes, the upper end of the mounting ear being square and the lower end being round.

[0014] By adopting the above technical solution, a unidirectional embedding structure with an ear and positioning structure that matches the shape can ensure accurate installation of the enzyme label strip and prevent it from being installed in reverse or shaking, thus ensuring the standardization of experimental operations and the accuracy of results.

[0015] Furthermore, the locking component is a locking plate that is rotatably connected to the side ear by a torsion spring, and the fastener is a bolt. When the locking plate is flipped over to cover, mechanical locking is achieved by screwing the bolt into the aligned fixing hole and connecting hole.

[0016] By adopting the above technical solution, the locking component is flipped over and covered, and then mechanically locked by screwing bolts into the fixing hole and the connecting hole. This can firmly fix the enzyme label strip, prevent it from loosening or popping out during the experiment, and ensure the safe and stable conduct of the experiment.

[0017] Furthermore, the edge height of the enzyme-labeled strip is higher than the inter-well junction, forming an isolation barrier.

[0018] By adopting the above technical solution, the edge height of the enzyme label strip well is higher than the connection between wells, forming an isolation barrier. This can prevent liquid overflow during sample addition or reaction, thus preventing cross-contamination and ensuring the purity and accuracy of experimental results.

[0019] Furthermore, the enzyme-labeled strips are made of polystyrene, and the plate holder is made of high-impact polystyrene, both of which meet biocompatibility standards.

[0020] By adopting the above technical solutions, the enzyme label strips are made of polystyrene and the plate holder is made of high-impact polystyrene, both of which meet biocompatibility standards. This reduces the impact on samples and reagents, ensuring the biosafety and reliability of the experiment.

[0021] In summary, the present invention has the following main advantages: 1. This utility model, by setting locking components, fixing holes and connecting holes, wherein the positioning structure at the top of the plate slot, such as the mounting slot, and the mounting ears on both sides of the enzyme label strip are shallowly mounted, realizes the initial positioning of the enzyme label strip, avoids excessive insertion and extraction force, and solves the problem of enzyme label strips being difficult to remove and easily broken due to excessive clamping in the prior art. Moreover, the locking components, such as locking plates, set on both sides of the top of the plate frame and connected by torsion springs, complete the final fixation. When it is necessary to remove the enzyme label strip, simply loosen the bolts, and the locking plates will automatically spring open under the action of the torsion spring, completely releasing the enzyme label strip, and the operator can easily remove it vertically from the positioning structure. 2. This utility model features a unidirectional embedding structure where the mounting ear and positioning structure are matched in shape. The upper end of the mounting ear is square and the lower end is round. When the enzyme label is placed into the slot of the enzyme label plate, this unique shape design ensures that the mounting ear can only be embedded into the positioning structure in a specific direction, effectively limiting the horizontal movement of the enzyme label. At the same time, the unidirectional embedding structure also prevents the enzyme label from easily detaching in the vertical direction. This solves the problem that the enzyme label may shake or shift in the slot of the enzyme label plate due to external factors, ensuring the stability of the enzyme label during reaction system construction, incubation, and other operations. It avoids affecting the accuracy and repeatability of experimental results due to shaking or shifting, and improves the quality of experimental data. Attached Figure Description

[0022] Figure 1 This is a three-dimensional structural diagram of the present invention; Figure 2 This is a partial three-dimensional structural diagram of the enzyme-labeled plate of this utility model; Figure 3 This is a partial three-dimensional structural diagram of the enzyme label strip of this utility model; Figure 4 This utility model Figure 1 Enlarged structural diagram at point A; Figure 5 This utility model Figure 2 A magnified structural diagram at point B in the middle.

[0023] In the diagram: 1. ELISA plate; 101. Plate holder; 102. Plate slot; 103. Insertion hole; 104. Positioning structure; 105. Fixing hole; 106. Side ear; 107. Locking component; 108. Connection hole; 2. ELISA strip; 201. Dropper; 202. Mounting ear. Detailed Implementation

[0024] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0025] Example 1: A V(D)J amplification optimization kit for immune repertoire sequencing, such as Figure 1-5 As shown, the device includes an enzyme-labeled plate 1 and an enzyme-labeled strip 2. The enzyme-labeled plate 1 includes a plate holder 101. The inner side of the plate holder 101 is provided with a plate groove 102 for carrying the enzyme-labeled strip 2. The upper part of the plate groove 102 is provided with a plurality of positioning structures 104 for positioning the enzyme-labeled strip 2. The top two sides of the plate holder 101 are provided with side ears 106. The side ears 106 are rotatably connected by torsion springs to locking components 107 for pressing and fixing the enzyme-labeled strip 2. The locking component 107 has a connecting hole 108, and the top of the plate holder 101 has a fixing hole 105 corresponding to the connecting hole 108. When the locking component 107 is flipped to the pressed state, the connecting hole 108 and the fixing hole 105 are aligned and locked by fasteners. The inner plate groove 102 can accommodate the enzyme label strip 2, providing a basic platform for the experiment. This design provides a stable place for experimental operations, ensures smooth experimental procedures, and improves experimental efficiency and the reliability of results.

[0026] Example 2: See Figure 1 , Figure 2 The enzyme-labeled strip 2 includes multiple dropper tubes 201, which are fixedly connected. Two mounting ears 202 are provided on each side of the row of dropper tubes 201. The positioning structure 104 is a mounting groove located on the upper inner wall of the plate groove 102. The mounting ears 202 are mounted in the mounting groove, and the multiple dropper tubes 201 are fixedly connected, facilitating batch sample addition. The mounting ears 202 on both sides can be mounted with the positioning structure 104 on the upper inner wall of the plate groove 102 of the enzyme-labeled plate 1, achieving stable fixation of the enzyme-labeled strip 2 on the enzyme-labeled plate 1, making experimental operations more convenient, reducing the risk of sample spillage, and ensuring the smooth progress of the experiment.

[0027] See Figure 1 , Figure 4 , Figure 5 The inner wall of the plate groove 102 has multiple insertion holes 103, which are used to insert multiple sets of enzyme-labeled strips 2 in a row. When multiple sets of enzyme-labeled strips 2 are inserted, the insertion holes 103 provide a stabilizing effect. This effectively prevents the enzyme-labeled strips 2 from shifting or moving during experimental operations and reactions, maintaining the stability of the experimental system and providing a strong guarantee for improving the accuracy of experimental results.

[0028] See Figure 1 , Figure 2 The dropper 201 of the enzyme labeling strip 2 features a flat-bottom design, and the well walls are surface-treated to enhance protein binding capacity. This flat-bottom design facilitates uniform sample distribution. Simultaneously, the surface-treated well walls enhance protein binding capacity, allowing proteins in the sample to better bind with reagents, promoting a complete reaction, improving reaction efficiency, and ultimately making experimental results more accurate and reliable, providing strong evidence for experimental analysis.

[0029] See Figure 3 , Figure 4 , Figure 5The mounting ear 202 and the positioning structure 104 are a shape-matched unidirectional embedding structure. The upper end of the mounting ear 202 is square and the lower end is round. This design ensures that the enzyme label strip 2 is accurately installed on the enzyme label plate 1, prevents reverse installation, and effectively limits the shaking of the enzyme label strip 2, ensuring standardized experimental operation and improving the accuracy of experimental results.

[0030] See Figure 1 , Figure 5 After the locking component 107 is flipped over and covered, it is mechanically locked by screwing the fixing hole 105 and the connecting hole 108. The locking component 107 is provided on the side ears 106 on both sides of the top of the plate holder 101 of the enzyme labeling plate 1 through a rotating shaft. When the locking component 107 is flipped over and covered, the connecting hole 108 on it is aligned with the fixing hole 105 at the front and rear ends of the top of the plate holder 101, and mechanical locking is achieved by screwing the bolts. This can firmly fix the enzyme labeling strip 2, prevent it from loosening or popping out during the experiment, and ensure the safety and stability of the experiment.

[0031] See Figure 1 , Figure 2 The edge of the well in enzyme label strip 2 is higher than the junction between wells, forming an isolation barrier. This effectively prevents liquid from overflowing from one dropper 201 into adjacent dropper 201 during sample addition or reaction, avoiding cross-contamination and ensuring the independence of the reaction environment for each sample, thereby guaranteeing the purity and accuracy of the experimental results.

[0032] See Figure 3 , Figure 4 , Figure 5 The enzyme label strip 2 is made of polystyrene, and the plate holder 101 of the enzyme label plate 1 is made of high-impact polystyrene. Both meet biocompatibility standards. The choice of materials can reduce chemical interference with samples and reagents, reduce the impact on biological activity, ensure the biosafety of the experiment, and provide a basis for obtaining reliable experimental results.

[0033] The implementation principle of this embodiment is as follows: First, when placing the enzyme label strip 2, the mounting ears 202 on both sides are embedded into the corresponding positioning structure 104 above the inner wall of the plate groove 102 of the enzyme label plate 1. Since the mounting ears 202 and the positioning structure 104 adopt a unidirectional embedding structure with matching shape, and the upper end of the mounting ears 202 is square and the lower end is round, this design can ensure that the enzyme label strip 2 is installed in the correct direction, effectively restrict its movement in the horizontal direction, and avoid displacement affecting the accuracy of the experiment. At the same time, the lower end of the enzyme label strip 2 is inserted into the insertion hole 103 provided below the inner wall of the plate groove 102 to further enhance its stability. Subsequently, the locking components 107 on the side ears 106 on both sides of the top of the flip plate holder 101 are connected by torsion springs and rotating shafts. The torsion springs provide a continuous reset torque, so that the locking components 107 automatically remain open when not locked, which facilitates the placement and removal of the enzyme label strip 2. When the locking components 107 are manually pressed down, the torque of the torsion springs provides an auxiliary pressing force to ensure that it stably covers the enzyme label strip 2. At this time, the connecting hole 108 on the locking components 107 is aligned with the fixing hole 105 opened on the top of the plate holder 101, and mechanical locking is achieved by screw bolts, thereby effectively preventing the enzyme label strip 2 from loosening or popping out during the experiment. During the experimental operation, the dropper 201 of the enzyme labeling strip 2 has a flat bottom design, and its well wall is surface treated to enhance protein binding ability, which helps the sample and reagent react fully. In addition, the edge of the well of the enzyme labeling strip 2 is higher than the connection between the wells, forming a physical isolation barrier to prevent liquid from overflowing during sample addition or reaction and avoid cross-contamination. The enzyme labeling strip 2 is made of polystyrene, and the plate holder 101 of the enzyme labeling plate 1 is made of high-impact polystyrene. Both meet the biocompatibility standards to ensure the safety and reliability of the experimental process. During the unlocking process, after the bolt is loosened and removed, the elastic potential energy stored in the torsion spring is immediately released, generating a reverse torque that drives the locking component 107 to automatically, quickly and smoothly flip open to the maximum opening angle and maintain a stable open state. This action completely removes the constraint on the enzyme label strip 2, providing the operator with an unobstructed space for removal. Due to the reset effect of the torsion spring, there is no need to manually flip or support the locking component, which not only simplifies the operation steps but also completely avoids interference or contamination that may be caused by additional operations, thus more effectively protecting the enzyme label strip 2 from breakage due to uneven force during the removal process. In addition, the number of dropper tubes 201 in the enzyme label strip 2 can be customized according to needs. The standard configuration is 12-20 wells. The number of wells shown in the figure is only for illustrating the design principle and is one of the embodiments. Moreover, the design of the ear 202 and the positioning structure 104 ensures that the enzyme label strip can be stably fixed regardless of the number of wells, avoiding experimental deviation and ensuring compatibility requirements.

[0034] Although embodiments of the present invention have been shown and described, these specific embodiments are merely explanations of the present invention and are not intended to limit the invention. The specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. After reading this specification, those skilled in the art may make modifications, substitutions, and variations to the embodiments as needed without departing from the principles and spirit of the present invention, provided that such modifications, substitutions, and variations are within the scope of the claims of the present invention and are protected by patent law.

Claims

1. A V(D)J amplification optimization kit for immune repertoire sequencing, characterized in that: The device includes an enzyme labeling plate (1) and an enzyme labeling strip (2). The enzyme labeling plate (1) includes a plate holder (101). The inner side of the plate holder (101) is provided with a plate groove (102) for carrying the enzyme labeling strip (2). The upper part of the plate groove (102) is provided with a plurality of positioning structures (104) for positioning the enzyme labeling strip (2). The top two sides of the plate holder (101) are provided with side ears (106). The side ears (106) are rotatably connected by a torsion spring to a locking component (107) for pressing and fixing the enzyme labeling strip (2). The locking component (107) has a connecting hole (108), and the top of the plate frame (101) has a fixing hole (105) corresponding to the connecting hole (108). When the locking component (107) is flipped to the pressing state, the connecting hole (108) and the fixing hole (105) are aligned and locked by fasteners.

2. The V(D)J amplification optimization kit for immune repertoire sequencing according to claim 1, characterized in that: The enzyme label strip (2) includes multiple drop tubes (201), which are fixedly connected to each other. Two mounting ears (202) are respectively provided on both sides of the drop tubes (201) in a row. The positioning structure (104) is a mounting groove opened on the upper part of the inner wall of the plate groove (102), and the mounting ears (202) are mounted with the mounting groove.

3. The V(D)J amplification optimization kit for immune repertoire sequencing according to claim 1, characterized in that: The inner wall of the plate groove (102) is provided with a plurality of insertion holes (103), which are used to insert multiple sets of enzyme label strips (2) in a row.

4. The V(D)J amplification optimization kit for immune repertoire sequencing according to claim 2, characterized in that: The dropper (201) has a flat bottom design and the pore walls are surface-treated to enhance protein binding ability.

5. The V(D)J amplification optimization kit for immune repertoire sequencing according to claim 2, characterized in that: The mounting ear (202) and the positioning structure (104) are a unidirectional embedded structure with matching shapes. The upper end of the mounting ear (202) is square and the lower end is round.

6. The V(D)J amplification optimization kit for immune repertoire sequencing according to claim 1, characterized in that: The locking component (107) is a locking piece that is rotatably connected to the side ear (106) by a torsion spring. The fastener is a bolt. When the locking piece is flipped over and covered, mechanical locking is achieved by screwing the bolt into the aligned fixing hole (105) and connecting hole (108).

7. The V(D)J amplification optimization kit for immune repertoire sequencing according to claim 1, characterized in that: The edge height of the well of the enzyme label strip (2) is higher than the inter-well connection, forming an isolation barrier.

8. The V(D)J amplification optimization kit for immune repertoire sequencing according to claim 1, characterized in that: The enzyme label strip (2) is made of polystyrene, and the plate holder (101) of the enzyme label plate (1) is made of high-impact polystyrene. Both meet the biocompatibility standards.

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